Skip to main content
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1974 Aug;71(8):3129–3133. doi: 10.1073/pnas.71.8.3129

Translation In Vitro of Total Nuclear RNA from HeLa Cell Nuclei Infected with Adenovirus 2

Nando K Chatterjee 1, Herbert Weissbach 1
PMCID: PMC388635  PMID: 4370520

Abstract

Total heterogeneous nuclear RNA from HeLa cells infected with adenovirus for 18-20 hr stimulates amino acid incorporation into protein in a cell-free system from Ehrlich ascites tumors. This stimulation of protein synthesis by the nuclear RNA requires intact nuclei. The role of nuclei in this system is unknown, but evidence is presented that the nuclei are involved in the conversion of high-molecular-weight RNA to low-molecular-weight species. Some of the newly synthesized polypeptides appear to resemble the virion polypeptides.

Keywords: nucleus, protein synthesis, DNA virus

Full text

PDF
3129

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Anderson C. W., Baum P. R., Gesteland R. F. Processing of adenovirus 2-induced proteins. J Virol. 1973 Aug;12(2):241–252. doi: 10.1128/jvi.12.2.241-252.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Berkowitz D. M., Kakefuda T., Sporn M. A simple and rapid method for the isolation of enzymatically active HeLa cell nuclei. J Cell Biol. 1969 Sep;42(3):851–854. doi: 10.1083/jcb.42.3.851. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brawerman G., Mendecki J., Lee S. Y. A procedure for the isolation of mammalian messenger ribonucleic acid. Biochemistry. 1972 Feb 15;11(4):637–641. doi: 10.1021/bi00754a027. [DOI] [PubMed] [Google Scholar]
  4. Brunner M., Raskas H. J. Processing of adenovirus RNA before release from isolated nuclei. Proc Natl Acad Sci U S A. 1972 Nov;69(11):3101–3104. doi: 10.1073/pnas.69.11.3101. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Caffier H., Green M. Adenovirus proteins. 3. Cell-free synthesis of adenovirus proteins in cytoplasmic extracts of KB cells. Virology. 1971 Oct;46(1):98–105. doi: 10.1016/0042-6822(71)90009-2. [DOI] [PubMed] [Google Scholar]
  6. Chatterjee N. K., Chuang D. M., Weissbach H. Further studies on eukaryote DNA stimulation of amino acid incorporation in E. coli extracts. Arch Biochem Biophys. 1974 Feb;160(2):603–613. doi: 10.1016/0003-9861(74)90437-8. [DOI] [PubMed] [Google Scholar]
  7. Chatterjee N. K., Kerwar S. S., Weissbach H. Initiation of protein synthesis in HeLa cells. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1375–1379. doi: 10.1073/pnas.69.6.1375. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chatterjee N. K., Weissbach H. Release of RNA from HeLa cell nuclei. Arch Biochem Biophys. 1973 Jul;157(1):160–167. doi: 10.1016/0003-9861(73)90401-3. [DOI] [PubMed] [Google Scholar]
  9. Darnell J. E., Jelinek W. R., Molloy G. R. Biogenesis of mRNA: genetic regulation in mammalian cells. Science. 1973 Sep 28;181(4106):1215–1221. doi: 10.1126/science.181.4106.1215. [DOI] [PubMed] [Google Scholar]
  10. Darnell J. E., Philipson L., Wall R., Adesnik M. Polyadenylic acid sequences: role in conversion of nuclear RNA into messenger RNA. Science. 1971 Oct 29;174(4008):507–510. doi: 10.1126/science.174.4008.507. [DOI] [PubMed] [Google Scholar]
  11. Darnell J. E., Wall R., Tushinski R. J. An adenylic acid-rich sequence in messenger RNA of HeLa cells and its possible relationship to reiterated sites in DNA. Proc Natl Acad Sci U S A. 1971 Jun;68(6):1321–1325. doi: 10.1073/pnas.68.6.1321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Edmonds M., Vaughan M. H., Jr, Nakazato H. Polyadenylic acid sequences in the heterogeneous nuclear RNA and rapidly-labeled polyribosomal RNA of HeLa cells: possible evidence for a precursor relationship. Proc Natl Acad Sci U S A. 1971 Jun;68(6):1336–1340. doi: 10.1073/pnas.68.6.1336. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Fujinaga K., Mak S., Green M. A method for determining the fraction of the viral genome transcribed during infection and its application to adenovirus-infected cells. Proc Natl Acad Sci U S A. 1968 Jul;60(3):959–966. doi: 10.1073/pnas.60.3.959. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Housman D., Pemberton R., Taber R. Synthesis of and chains of rabbit hemoglobin in a cell-free extract from Krebs II ascites cells. Proc Natl Acad Sci U S A. 1971 Nov;68(11):2716–2719. doi: 10.1073/pnas.68.11.2716. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  16. Lee S. Y., Mendecki J., Brawerman G. A polynucleotide segment rich in adenylic acid in the rapidly-labeled polyribosomal RNA component of mouse sarcoma 180 ascites cells. Proc Natl Acad Sci U S A. 1971 Jun;68(6):1331–1335. doi: 10.1073/pnas.68.6.1331. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lindberg U., Darnell J. E. SV40-specific RNA in the nucleus and polyribosomes of transformed cells. Proc Natl Acad Sci U S A. 1970 Apr;65(4):1089–1096. doi: 10.1073/pnas.65.4.1089. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Maizel J. V., Jr, White D. O., Scharff M. D. The polypeptides of adenovirus. I. Evidence for multiple protein components in the virion and a comparison of types 2, 7A, and 12. Virology. 1968 Sep;36(1):115–125. doi: 10.1016/0042-6822(68)90121-9. [DOI] [PubMed] [Google Scholar]
  19. Parsons J. T., Gardner J., Green M. Biochemical studies on adenovirus multiplication, XIX. Resolution of late viral RNA species in the nucleus and cytoplasm. Proc Natl Acad Sci U S A. 1971 Mar;68(3):557–560. doi: 10.1073/pnas.68.3.557. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Parsons J. T., Green M. Biochemical studies on adenovirus multiplication. 18. Resolution of early virus-specific RNA species in Ad 2 infected and transformed cells. Virology. 1971 Jul;45(1):154–162. doi: 10.1016/0042-6822(71)90122-x. [DOI] [PubMed] [Google Scholar]
  21. Penman S., Vesco C., Penman M. Localization and kinetics of formation of nuclear heterodisperse RNA, cytoplasmic heterodisperse RNA and polyribosome-associated messenger RNA in HeLa cells. J Mol Biol. 1968 May 28;34(1):49–60. doi: 10.1016/0022-2836(68)90234-9. [DOI] [PubMed] [Google Scholar]
  22. Ruiz-Carrilo A., Beato M., Schutz G., Feigelson P., Allfrey V. G. Cell-free translation of the globin message within polydisperse high-molecular-weight ribonucleic acid of avian erythrocytes. Proc Natl Acad Sci U S A. 1973 Dec;70(12):3641–3645. doi: 10.1073/pnas.70.12.3641. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Scherrer K., Marcaud L., Zajdela F., London I. M., Gros F. Patterns of RNA metabolism in a differentiated cell: a rapidly labeled, unstable 60S RNA with messenger properties in duck erythroblasts. Proc Natl Acad Sci U S A. 1966 Nov;56(5):1571–1578. doi: 10.1073/pnas.56.5.1571. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Soeiro R., Vaughan M. H., Warner J. R., Darnell J. E., Jr The turnover of nuclear DNA-like RNA in HeLa cells. J Cell Biol. 1968 Oct;39(1):112–118. doi: 10.1083/jcb.39.1.112. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Stevens R. H., Williamson A. R. Specific IgG mRNA molecules from myeloma cells in heterogeneous nuclear and cytoplasmic RNA containing poly-A. Nature. 1972 Sep 15;239(5368):143–146. doi: 10.1038/239143a0. [DOI] [PubMed] [Google Scholar]
  26. Wall R., Philipson L., Darnell J. E. Processing of adenovirus specific nuclear RNA during virus replication. Virology. 1972 Oct;50(1):27–34. doi: 10.1016/0042-6822(72)90342-x. [DOI] [PubMed] [Google Scholar]
  27. Williamson R., Drewienkiewicz C. E., Paul J. Globin messenger sequences in high molecular weight RNA from embryonic mouse liver. Nat New Biol. 1973 Jan 17;241(107):66–68. doi: 10.1038/newbio241066a0. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES